BACILLUS AS AQUACULTURE PROBIOTICS: POTENTIAL AND LIMITATIONS
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1 BACILLUS AS AQUACULTURE PROBIOTICS: POTENTIAL AND LIMITATIONS O. Decamp INVE TECHNOLOGIES nv, Belgium. D.J.W. Moriarty The University of Queensland, Australia, Thailand.
2 Bacillus as probiotic for animal use No authorised additive EU Many products authorised Asia /Latin America Bacillus cereus var. toyoi Bacillus coagulans Bacillus lentus Bacillus subtilis Bacillus pumilus Bacillus licheniformis Authorised additive EU Bacillus cereus var. toyoi (Toyocerin) Bacillus subtilis (Belfeed, Calsporin, Bioplus 2B) Bacillus licheniformis (Bioplus 2B) ex_fr.html
3 Benefits of selected Bacillus strains Present in aquatic animals and environments Ability to grow under wide range of conditions Producer of numerous enzymes Active against many pathogens Directly by growth inhibition Indirectly by competitive exclusion Advantages of spores Long shelf life Easy to transport Resistant to dehydration Resistant to temperatures up to 60 C
4 PROBIOTICS AS ALTERNATIVE TO PROPHYLACTIC USE OF ANTIMICROBIALS: LIMITATIONS AND POTENTIAL Doubts of efficacy Doubts of composition Doubts of safety Suitable claims Suitable delivery Suitable QC Suitable production Suitable registration
5 Advantages of Bacillus mixtures Doubts of composition safety Germination not fermentation Bacillus Spores high concentration Freeze-dried or liquid culture low concentration <8 hours for activation germination 24 hours for fermentation contamination
6 PROBIOTICS AS ALTERNATIVE TO PROPHYLACTIC USE OF ANTIMICROBIALS: LIMITATIONS AND POTENTIAL R&D Registration Safety to environment Farmers Safety to target Formulations COST/PRACTICALITIES Screening of strains for function
7 Function: from lab to pilot-scale Lab Pathogen control Field test Sanolife MIC
8 SAFETY: environment including humans Non-target aquaculture organisms
9 SAFETY: Survival of starved seabream larvae 100 Survival of seabream larvae (%) A 3 LMG i8 10i isol anm Days after hatching
10 SAFETY: Survival of starved seabream larvae Bacillus mixture conc1 Bacillus mixture conc2 Filtered w ater Sterile w ater Survival (%) DAH
11 SAFETY: environment including humans Humans Bacillus cereus B. cereus enterotoxins emetic toxin genes
12 PROBIOTICS AS ALTERNATIVE TO PROPHYLACTIC USE OF ANTIMICROBIALS: LIMITATIONS AND POTENTIAL Commercial Sales Registration QC Support R&D Upscale production COST? Safety issues
13 UPSCALE PRODUCTION Advantage of Bacillus
14 UPSCALE PRODUCTION? Species with little history Cost!!!
15 Feed back from field
16 Shrimp Grow-Out Probiotics Sanolife PRO-1/PRO-2 mixed with feed (min 10 8 cfu/g feed) $ $ US Dollars $2.000 $1.600 $1.200 $ Indian Rupees $ $0 Controls 5-7 Treated 2,3 & 4 Treated 2 & 3 0
17 Alternative delivery incorporation in feed Advantages of Bacillus Bacillus spores can exist in a desiccated and dehydrated state almost indefinitely. The resistance of spores can be explained by various factors: - thick proteinaceous spore coats - reduced permeability of the spore core - reduced water content - etc (Setlow 1999)
18 Alternative delivery incorporation in feed The extrusion-expansion and drying process resulted in the loss of >99 % of spores Biourge et al The use of probiotics in the diet of dogs. J Nutr Dec;128(12 Suppl):2730S-2732S. Selection of resistant strains What about claims? Ruiz et al. J. Food Sci. 67(2)
19 -20 C 4 C Room 30 C Alternative delivery incorporation in feed 1. Loss during production 2. Loss during storage 1 month storage Sanolife PRO-1 temperature 1,E+08 1,E+07 1,E+06 1,E+05 Spore concentration (10^6 cfu/g) 1,8 Batch 1 1,6 Batch 2 1,4 Batch 3 Batch 4 1,2 1,0 0,8 0,6 0,4 0,2 0, Storage time (month) Biourge et al J. Nutrition 128 1,E+04 1,E+03
20 FURTHER INVESTIGATION -Legal status -Spore germination, proliferation, colonisation - Resistant pathogens (geographical variations, history, management, etc) - Follow-up of probiotics
21 Thank you
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